期刊
MICROBIAL CELL FACTORIES
卷 19, 期 1, 页码 -出版社
BMC
DOI: 10.1186/s12934-020-01415-z
关键词
Nicotinamide mononucleotide; Noncanonical redox cofactor; Escherichia coli; Metabolic engineering; NAD(+)biosynthesis; Biomimetic cofactor
资金
- University of California, Irvine
- National Science Foundation (NSF) [1847705]
- National Institutes of Health (NIH) [DP2 GM137427]
- Graduate Assistance in Areas of National Need fellowship - U.S. Department of Education
- Federal Work Study Program - U.S. Department of Education
- Div Of Chem, Bioeng, Env, & Transp Sys
- Directorate For Engineering [1847705] Funding Source: National Science Foundation
Background Noncanonical redox cofactors are emerging as important tools in cell-free biosynthesis to increase the economic viability, to enable exquisite control, and to expand the range of chemistries accessible. However, these noncanonical redox cofactors need to be biologically synthesized to achieve full integration with renewable biomanufacturing processes. Results In this work, we engineeredEscherichia colicells to biosynthesize the noncanonical cofactor nicotinamide mononucleotide (NMN+), which has been efficiently used in cell-free biosynthesis. First, we developed a growth-based screening platform to identify effective NMN(+)biosynthetic pathways inE. coli. Second, we explored various pathway combinations and host gene disruption to achieve an intracellular level of similar to 1.5 mM NMN+, a 130-fold increase over the cell's basal level, in the best strain, which features a previously uncharacterized nicotinamide phosphoribosyltransferase (NadV) fromRalstonia solanacearum.Last, we revealed mechanisms through which NMN(+)accumulation impactsE. colicell fitness, which sheds light on future work aiming to improve the production of this noncanonical redox cofactor. Conclusion These results further the understanding of effective production and integration of NMN(+)intoE. coli. This may enable the implementation of NMN+-directed biocatalysis without the need for exogenous cofactor supply.
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